scholarly journals Peptide translocation by the lysosomal ABC transporter TAPL is regulated by coupling efficiency and activation energy

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Christoph Bock ◽  
Tina Zollmann ◽  
Katharina-Astrid Lindt ◽  
Robert Tampé ◽  
Rupert Abele
2015 ◽  
Vol 112 (7) ◽  
pp. 2046-2051 ◽  
Author(s):  
Tina Zollmann ◽  
Gemma Moiset ◽  
Franz Tumulka ◽  
Robert Tampé ◽  
Bert Poolman ◽  
...  

ATP-binding cassette (ABC) transporters use ATP to drive solute transport across biological membranes. Members of this superfamily have crucial roles in cell physiology, and some of the transporters are linked to severe diseases. However, understanding of the transport mechanism, especially of human ABC exporters, is scarce. We reconstituted the human lysosomal polypeptide ABC transporter TAPL, expressed inPichia pastoris, into lipid vesicles (liposomes) and performed explicit transport measurements. We analyzed solute transport at the single liposome level by monitoring the coincident fluorescence of solutes and proteoliposomes in the focal volume of a confocal microscope. We determined a turnover number of eight peptides per minute, which is two orders of magnitude higher than previously estimated from macroscopic measurements. Moreover, we show that TAPL translocates peptides against a large concentration gradient. Maximal filling is not limited by an electrochemical gradient but by trans-inhibition. Countertransport and reversibility studies demonstrate that peptide translocation is a strictly unidirectional process. Altogether, these data are included in a refined model of solute transport by ABC exporters.


2018 ◽  
Vol 430 (6) ◽  
pp. 853-866 ◽  
Author(s):  
Jelger A. Lycklama a Nijeholt ◽  
Ruslan Vietrov ◽  
Gea K. Schuurman-Wolters ◽  
Bert Poolman

2019 ◽  
Author(s):  
Chengcheng Fan ◽  
Jens T. Kaiser ◽  
Douglas C. Rees

AbstractATP Binding Cassette (ABC) transporters often exhibit significant basal ATPase activity in the absence of transported substrates. To investigate the factors that contribute to this inefficient coupling of ATP hydrolysis to transport, we characterized the structures and functions of variants of the bacterial Atm1 homolog from Novosphingobium aromaticivorans (NaAtm1), including forms with disulfide crosslinks between the nucleotide binding domains. Unexpectedly, disulfide crosslinked variants of NaAtm1 reconstituted into proteoliposomes not only transported oxidized glutathione, but also exhibited more efficient coupling of ATP hydrolysis to GSSG transport than the native transporter. These observations suggest that enhanced conformational dynamics of reconstituted NaAtm1 may contribute to the inefficient use of ATP. Understanding the origins of this uncoupled ATPase activity, and reducing the impact through disulfide crosslinking or other protocols, will be critical for the detailed dissection of ABC transporter mechanism to assure that the ATP dependent steps are indeed relevant to substrate translocation.


2017 ◽  
Vol 77 (04) ◽  
pp. 379-395
Author(s):  
L Schmieding ◽  
A Klein ◽  
N Maass ◽  
C Eckmann-Scholz ◽  
D Lütjohann ◽  
...  

2007 ◽  
Vol 34 (S 2) ◽  
Author(s):  
S Cotte ◽  
N Kruse ◽  
N Ahsen ◽  
UK Zettl ◽  
R Gold ◽  
...  
Keyword(s):  

2003 ◽  
Vol 762 ◽  
Author(s):  
A. Gordijn ◽  
J.K. Rath ◽  
R.E.I. Schropp

AbstractDue to the high temperatures used for high deposition rate microcrystalline (μc-Si:H) and polycrystalline silicon, there is a need for compact and temperature-stable doped layers. In this study we report on films grown by the layer-by-layer method (LbL) using VHF PECVD. Growth of an amorphous silicon layer is alternated by a hydrogen plasma treatment. In LbL, the surface reactions are separated time-wise from the nucleation in the bulk. We observed that it is possible to incorporate dopant atoms in the layer, without disturbing the nucleation. Even at high substrate temperatures (up to 400°C) doped layers can be made microcrystalline. At these temperatures, in the continuous wave case, crystallinity is hindered, which is generally attributed to the out-diffusion of hydrogen from the surface and the presence of impurities (dopants).We observe that the parameter window for the treatment time for p-layers is smaller compared to n-layers. Moreover we observe that for high temperatures, the nucleation of p-layers is more adversely affected than for n-layers. Thin, doped layers have been structurally, optically and electrically characterized. The best n-layer made at 400°C, with a thickness of only 31 nm, had an activation energy of 0.056 eV and a dark conductivity of 2.7 S/cm, while the best p-layer made at 350°C, with a thickness of 29 nm, had an activation energy of 0.11 V and a dark conductivity of 0.1 S/cm. The suitability of these high temperature n-layers has been demonstrated in an n-i-p microcrystalline silicon solar cell with an unoptimized μc-Si:H i-layer deposited at 250°C and without buffer. The Voc of the cell is 0.48 V and the fill factor is 70 %.


Diabetes ◽  
2018 ◽  
Vol 67 (Supplement 1) ◽  
pp. 1897-P
Author(s):  
HISASHI YOKOMIZO ◽  
ATSUSHI ISHIKADO ◽  
TAKANORI SHINJO ◽  
KYOUNGMIN PARK ◽  
YASUTAKA MAEDA ◽  
...  

Author(s):  
А. А. Горват ◽  
В. М. Кришеник ◽  
А. Е. Кріштофорій ◽  
В. В. Мінькович ◽  
О. А. Молнар

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